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Application of india ink to the peritoneal and pleural surfaces of the adult human diaphragm allowed visualization of the distribution and morphology of the lymphatic vessels by light microscopy and scanning electron microscopy. The diaphragms examined had been fixed and stored in 10% formalin. Numerous lymphatic vessels were stained black with india ink, presenting reticular, radial-meshwork, ladder-like and lacy patterns. They were distributed throughout the entire sternocostal part. Analysis by light and scanning electron microscopy of the areas indicated by india ink revealed the presence of primary lymphatic vessels that formed lymphatic lacunae and stomatal openings to the peritoneal cavity. A layer of secondary collecting lymphatic vessels was located cranially with respect to the layer of primary lymphatic vessels. Thus, the peritoneum had at least two layers of lymphatic vessels. These lymphatic vessels were not tubular vessels but resembled flat cisternae, as has been suggested in the case of the mouse diaphragm. The pleura lacked lymphatic stomata and had no such double-layered lymphatic organization. This is the first report that showed distribution and morphology of the lymphatic vessels in the diaphragmatic peritoneum of the formalin-fixed, adult human diaphragm. The method and results in the present study may contribute to morphological analysis of the lymphatic system in the wall of the human body cavity.

The Distribution and Morphology of Lymphatic Vessels on the Peritoneal Surface of the Adult Human Diaphragm, as Revealed by an Ink-absorption Method